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Cytogenetic Analysis of Tegenaria Elysii (Araneae: Agelenidae)

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Cytogenetic Analysis of Tegenaria Elysii (Araneae: Agelenidae) Sakarya University Journal of Science, 22 (6), 1669-1673, 2018. SAKARYA UNIVERSITY JOURNAL OF SCIENCE e-ISSN: 2147-835X http://www.saujs.sakarya.edu.tr Receved 07-12-2017 Do Accepted 10.16984/saufenblder.363843 17-04-2018 Cytogenetic Analysis of Tegenaria elysii (Araneae: Agelenidae) Ümit KUMBIÇAK*1 Abstract There are 61 species of agelenid spiders in our country and the genus Tegenaria represents the widest group in terms of species number. In this study, it was aimed to investigate the cytogenetic properties of Tegenaria elysii. Chromosomes were obtained by making some modifications to the method of Pekár and Král (2001). Hypotonic, fixation and staining steps were applied to the gonads respectively. As a result, the number of diploid chromosomes and the sex chromosome system were found as 2n=42, ♂X1X20/♀X1X1X2X2. Total relative lengths of chromosomes decreased gradually from 6,83% to 3,31%. Sex chromosomes which positive heteropycnotics in the stage of meiosis. I have been detected to be isopycnic in the stage of meiosis II. 20 autosomal bivalent and two sex chromosomes were identified in which diplotene and diakinesis stages. Keywords: Agelenidae, chromosome, karyotype to the subclass Araneomorphae and easily 1. INTRODUCTION distinguished from other families by its characteristics posterior spinneret which has two- segments and long, thin, tipped toward each other. Spiders are one of the largest animal group, and Tegenaria Latreille 1804 differs from other genera contain almost 47 000 species all around the world by the number and position of gonadal teeth, the [1]. In our country, a total of 1117 spider species structure of the tibial and median apophyses, the belonging to 52 families were determined [2]. shape of the conductor, the shape of the vulva only Until now, although studies have been carried out having a spiral duct, and the scleroid structure [5]. in systematics, fauna and ecology, the cytogenetic The araneomorph karyotypes are characterized by investigations is scarce. The fact that tiny a predominance of acrocentric chromosomes, chromosome morphology, the presence of X1X20 sex chromosome system [6], relatively low different sex chromosome systems and the diploid chromosome numbers (ranges from 10 to requirement of modification in the method for 49, [7]), and chiasmatic meiosis. each species limits the investigations on spider Acrocentric/telocentric karyotypes of entelegynes cytogenetics [3]. with lower chromosome numbers could be derived Spiders are divided into three subclasses namely from ancestral karyotypes by tandem fusions [8] or Mesothelae, Mygalomorphae and Araneomorphae by cycles of centric fusions and subsequent [4]. Among them, Agelenidae family is belonging pericentric inversions [7]. The latter hypothesis is * Corresponding Author 1 Nevşehir Hacı Bektaş Veli University, Faculty of Art and Sciences, Department of Molecular Biology and Genetics, Nevşehir, Turkey- [email protected] 1 1669 "Ümit Kumbıçak Cytogenetic analysis of tegenaria elysii (araneae: agelenidae)…" supported by the fact that centric fusions are the acid solution on heating plate that surface most frequent source of chromosome temperature 42 °C (for 20 min). The slides were polymorphism found in populations of entelegyne air dried for overnight and stained with 5% Giemsa spiders [7, 9]. In this study, karyotype solution in Sorensen phosphate buffer (ph=6.8) for characteristics and meiotic behavior of Tegenaria 50 min. Chromosome slides were investigated an elysii Brignoli, 1978 belonging to Agelenidae BX53 microscope (Olympus) and well spread family, which is found in our country, were stages were photographed using DP26 digital investigated for the first time. camera with CellSens software (Olympus). The diploid chromosome number were determined by 2. EXPERIMENTAL the mitotic metaphases or diplotene stages. Relative chromosome lengths (RCL) including standard deviations were calculated as a The male specimens of Tegenaria elysii were percentage of the total chromosome length of the collected from Nevşehir, Kayseri and diploid set by CellSens software. Classification of Kahramanmaraş between March to May in the chromosome morphology was made by the year 2015 from different habitats and localities. protocol of (Table 2) [11]. The collection were made by hands or pitfall traps (Table 1). During the field study, no treatment was Tablo 2. Determination of chromosome morphology applied to the spiders, and each one was taken to according to the centromere position (C) and arm ratio (q/p) falcon tubes and placed in the laboratory. Male gonads were used in obtaining chromosomes due Centromere Arm ratio Chromosome to the lots of cells contained. Therefore, it was position (q/p) morphology possible to determine various mitotic and meiotic stages from males. However, females were not Median 1.00-1.70 Metacentric used because of the low division frequency. Submedian 1.71-3.00 Submetacentric Table 1. Data of collection date, locality information and Subterminal 3.01-7.00 Subtelocentric number of specimens used in this study Terminal 7.01-∞ Acrocentric Collection Number Localities Date of species Region Coordinates 3. RESULTS Nevşehir, 38°32'24.58''N 14.03.2015 2♂♂ Acıgöl 34°32'51.72''E In this study, cytogenetic structure, diploid Kayseri, 38°42'47.93''N 11.04.2015 1♂ Pınarbaşı 36°22'49.75''E number, sex chromosome system and meiosis Kahramanma- 38°00'42.27''N characteristics of Tegenaria elysii Brignoli, 1978 19.04.2015 3♂♂ raş, Göksun 36°28'15.80''E were determined for the first time. Kayseri, 38°42'45.51''N 23.05.2015 7♂♂ Pınarbaşı 36°23'26.84''E 1.1. Karyotype and sex chromosome system of T. elysii The chromosome preparations were performed according to the method of [10] with some The chromosome set of male T. elysii (2n=42, modifications. Alive male specimens were X1X20) contained 42 chromosomes with squeezed from the prosoma and separated from the telocentric morphology (Fig.1, 2). opistosoma. Prosoma and pedipalps were kept in 70% ethanol for systematic diagnosis. The gonads were dissected out in physiological solution for invertebrates. Then three steps were used; hypotonic solution in 0.075 M KCl (for 15 min), twice fixation in methanol:glacial acetic acid (3:1 for 10 and 20 min); spreading in 60% glacial acetic 2 Sakarya University Journal of Science, 22 (6), 1669-1673, 2018. 1670 "Ümit Kumbıçak Cytogenetic analysis of tegenaria elysii (araneae: agelenidae)…" Table 3. Chromosome length measurements of T. elysii: short arm (p); long arm (q); relative lengths (p+q); arm ratio (q/p); TCL: total chromosome lengths (%); CM: chromosome morphology; T: telocentric ; ± standart deviation) p q (p+q) q/p TCL µm µm (%) CM 1 0 10,88±0,67 10,88±0,67 ∞ 6,83 T 2 0 9,46±0,4 9,46±0,4 ∞ 5,93 T 3 0 8,77±0,37 8,77±0,37 ∞ 5,5 T 4 0 8,32±0,36 8,32±0,36 ∞ 5,22 T Fig.1. Karyotype of T. elysii based on spermatogonial 5 0 8,07±0,4 8,07±0,4 ∞ 5,06 T metaphases (Scale=10 µm) 6 0 7,87±0,43 7,87±0,43 ∞ 4,94 T 7 0 7,69±0,33 7,69±0,33 ∞ 4,82 T Autosome pairs decreased gradually in size from 8 0 7,51±0,35 7,51±0,35 ∞ 4,71 T 6,83% to 3,31% of total chromosome length 9 0 7,37±0,29 7,37±0,29 ∞ 4,62 T (TCL). Relative length of X1 and X2 were 4,24% 10 0 7,22±0,32 7,22±0,32 ∞ 4,53 T and 3,92% of TCL, respectively (Table 3). X1 was longer than the 14th autosome pair and X2 was 11 0 7,08±0,28 7,08±0,28 ∞ 4,44 T longer than the 16th autosomal pair. X1 and X2 12 0 6,97±0,25 6,97±0,25 ∞ 4,37 T were in similar size. 13 0 6,81±0,3 6,81±0,3 ∞ 4,27 T 14 0 6,66±0,28 6,66±0,28 ∞ 4,18 T 15 0 6,45±0,23 6,45±0,23 ∞ 4,05 T 16 0 6,19±0,42 6,19±0,42 ∞ 3,88 T 17 0 6,13±0,44 6,13±0,44 ∞ 3,85 T 18 0 5,93±0,5 5,93±0,5 ∞ 3,72 T 19 0 5,76±0,55 5,76±0,55 ∞ 3,61 T 20 0 5,27±0,62 5,27±0,62 ∞ 3,31 T X1 0 6,76±0,35 6,76±0,35 ∞ 4,24 T X2 0 6,25±0,39 6,25±0,39 ∞ 3,92 T Fig. 2. Idiogram of T. elysii based on the haploid set of chromosomes 1.2. Meiotic characteristics of T. elysii The sex chromosomes were stained positively heteropycnotic during the first substages of prophase I (i.e. leptotene, zygotene and pacythene) (Fig. 3.a). 20 autosomal biavelents and two univalent sex chromosomes were determined in diplotene, diakinesis and metaphase I. The bivalents had one chiasma (or sometimes two chiasmata) with terminal, interstitial and proximal type (Fig. 3.b). All chromosomes including sex chromosomes were “V” shaped in anaphase I and sex chromosomes were located on the periphery of nucleus (Fig. 3.c). During the second meiotic stages (i.e. prophase II, metaphase II and anaphase II), the sex chromosomes were isopycnotic but 3 Sakarya University Journal of Science, 22 (6), 1669-1673, 2018. 1671 "Ümit Kumbıçak Cytogenetic analysis of tegenaria elysii (araneae: agelenidae)…" were easily distinguished because of their earlier Strand, 1906); Tegenaria campestris (C.L. Koch, condensation (Fig. 3.d). 1834); Tegenaria domestica (Clerck, 1757); Tegenaria ferruginea (Panzer, 1804); Tegenaria parietina (Fourcroy, 1785) ve Tegenaria silvestris L. Koch, 1872 [12]. According to these previously studies, the diploid chromosome number were determined between 2n♂=18 (E.
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  • Hobo Spider, Eratigena Agrestis, Is a Fig
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  • Phylogeny of Entelegyne Spiders: Affinities of the Family Penestomidae
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